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Chlamydia psittaci PmpD-N Exacerbated Chicken Macrophage Function by Triggering Th2 Polarization and the TLR2/MyD88/NF-kappa B Signaling Pathway

文献类型： 外文期刊

作者： Chu, Jun 1 ; Li, Xiaohui 1 ; Qu, Guanggang 3 ; Wang, Yihui 1 ; Li, Qiang 1 ; Guo, Yongxia 1 ; Hou, Lei 2 ; Liu, Jue 2 ; Eko, 1 ;

作者机构： 1.China Agr Univ, Coll Vet Med, Key Lab Anim Epidemiol & Zoonosis, Minist Agr, Beijing 100193, Peoples R China

2.Beijing Acad Agr & Forestry Sci, Inst Anim Husb & Vet Med, Beijing Key Lab Prevent & Control Infect Dis Live, Beijing 100097, Peoples R China

3.Shandong Binzhou Anim Sci & Vet Med Acad, Binzhou 256600, Peoples R China

4.Morehouse Sch Med, Dept Microbiol Biochem & Immunol, Atlanta, GA 30310 USA

关键词： Chlamydia psittaci; HD11 macrophage; PmpD-N; signaling pathway; Th2 immune response

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）

ISSN：

年卷期： 2020 年 21 卷 6 期

页码：

收录情况： SCI

摘要： The polymorphic membrane protein D (PmpD) is a highly conserved outer membrane protein which plays an important role in pathogenesis during Chlamydia psittaci infection. In this study, we evaluated the ability of the N-terminus of PmpD (PmpD-N) to modulate the functions of chicken macrophages and the signaling pathway(s) involved in PmpD-N-induced Toll-like receptors (TLRs), as well as interleukin (IL)-6 and IL-10 cytokine secretions. Thus, HD11 macrophages were treated with exogenous and intracellular PmpD-N of C. psittaci. The chlamydial growth was evaluated by enumeration of chlamydial loads in the infected macrophages. The phagocytic function of macrophages following PmpD-N treatment was detected by fluorescein-labeled Escherichia coli (E. coli). The concentration of nitric oxide (NO) secreted by HD11 macrophages was measured by the amount of NO2- in the culture supernatant using the Griess method. The cytokine secretions were assessed using multiplex cytokine ELISA kits. Expression levels of TLRs, myeloid differentiation factor 88 (MyD88), and nuclear factor kappa B (NF-kappa B) were analyzed by a Western blotting assay, as well as a luciferase assay, while NF-kappa B p65 nuclear translocation was assessed by confocal microscopy. The nuclear translocation of the transcription factor NF-kappa B was confirmed by evaluating its ability to combine with the corresponding promoter using the electrophoretic mobility shift assay (EMSA). After treatment with exogenous or endogenous PmpD-N, chlamydial loads and phagocytic functions were reduced significantly compared with those of the plasmid vector group, while NO secretions were reduced significantly compared with those of the lipopolysaccharide (LPS) treatment. Stimulation of HD11 cells with PmpD-N provoked the secretion of the Th2 cytokines, IL-6, and IL-10 and upregulated the expression of TLR2, TLR4, MyD88, and NF-kappa B. Furthermore, inhibition of TLR2, MyD88, and NF-kappa B in HD11 cells significantly decreased IL-6 and IL-10 cytokine levels, while NO production and phagocytosis increased significantly, strongly suggesting their involvement in PmpD-N-induced Th2 cytokine secretion and macrophage dysfunction. Our data indicate that C. psittaci PmpD-N inhibited macrophage functions by activating the Th2 immune response and the TLR2/MyD88/NF-kappa B signaling pathway.

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